Collateral Health Issues Derived from the Covid-19 Pandemic.

At the end of 2019, a new coronavirus (Covid-19) outbreak occurred in Wuhan, China, and spread throughout the world despite efforts to contain the virus. At the end of January 2020, the General Director of the World Health Organization (WHO) declared a Public Health Emergency of International Concern, and by mid-May 2020, the worldwide number of known Covid-19 cases had surpassed 4.4 million including more than 300,000 deaths..

Histone H2A and H2B Are Monoubiquitinated at AID-Targeted Loci

Background: Somatic hypermutation introduces base substitutions into the rearranged and expressed immunoglobulin (Ig) variable regions to promote immunity. This pathway requires and is initiated by the Activation Induced Deaminase (AID) protein, which deaminates cytidine to produce uracils and UG mismatches at the Ig genes. Subsequent processing of uracil by mismatch repair and base excision repair factors contributes to mutagenesis. While selective for certain genomic targets, the chromatin modifications which distinguish hypermutating from non-hypermutating loci are not defined. Methodology/Principal Findings: Here, we show that AID-targeted loci in mammalian B cells contain ubiquitinated chromatin. Chromatin immunoprecipitation (ChIP) analysis of a constitutively hypermutating Burkitt\u27s B cell line, Ramos, revealed the presence of monoubiquitinated forms of both histone H2A and H2B at two AID-associated loci, but not at control loci which are expressed but not hypermutated. Similar analysis using LPS activated primary murine splenocytes showed enrichment of the expressed V(H) and S gamma 3 switch regions upon ChIP with antibody specific to AID and to monoubiquitinated H2A and H2B. In the mechanism of mammalian hypermutation, AID may interact with ubiquitinated chromatin because confocal immunofluorescence microscopy visualized AID colocalized with monoubiquitinated H2B within discrete nuclear foci. Conclusions/Significance: Our results indicate that monoubiquitinated histones accompany active somatic hypermutation, revealing part of the histone code marking AID-targeted loci. This expands the current view of the chromatin state during hypermutation by identifying a specific nucleosome architecture associated with somatic hypermutation

Genome Instability and Transcription Elongation Impairment in Human Cells Depleted of THO/TREX

THO/TREX connects transcription with genome integrity in yeast, but a role of mammalian THO in these processes is uncertain, which suggests a differential implication of mRNP biogenesis factors in genome integrity in yeast and humans. We show that human THO depletion impairs transcription elongation and mRNA export and increases instability associated with DNA breaks, leading to hyper-recombination and γH2AX and 53BP1 foci accumulation. This is accompanied by replication alteration as determined by DNA combing. Genome instability is R-loop–dependent, as deduced from the ability of the AID enzyme to increase DNA damage and of RNaseH to reduce it, or from the enhancement of R-loop–dependent class-switching caused by THOC1-depletion in CH12 murine cells. Therefore, mammalian THO prevents R-loop formation and has a role in genome dynamics and function consistent with an evolutionary conservation of the functional connection between these mRNP biogenesis factors and genome integrity that had not been anticipated

Negative Supercoiling Creates Single-Stranded Patches of DNA That Are Substrates for AID–Mediated Mutagenesis

Antibody diversification necessitates targeted mutation of regions within the immunoglobulin locus by activation-induced cytidine deaminase (AID). While AID is known to act on single-stranded DNA (ssDNA), the source, structure, and distribution of these substrates in vivo remain unclear. Using the technique of in situ bisulfite treatment, we characterized these substrates—which we found to be unique to actively transcribed genes—as short ssDNA regions, that are equally distributed on both DNA strands. We found that the frequencies of these ssDNA patches act as accurate predictors of AID activity at reporter genes in hypermutating and class switching B cells as well as in Escherichia coli. Importantly, these ssDNA patches rely on transcription, and we report that transcription-induced negative supercoiling enhances both ssDNA tract formation and AID mutagenesis. In addition, RNaseH1 expression does not impact the formation of these ssDNA tracts indicating that these structures are distinct from R-loops. These data emphasize the notion that these transcription-generated ssDNA tracts are one of many in vivo substrates for AID

Validation of a hand-held lactate device in determination of blood lactate in critically injured patients

OBJECTIVESAdmission blood lactate is an accurate predictor of injury severity and mortality in trauma patients. The purpose of this study was to evaluate a portable lactate analyzer in a clinical setting by patient care staff. DESIGNA prospective, single-operator control solution and patient sample study, using two test devices and a reference device. SETTINGAn urban Level I trauma center. PATIENTSA convenience sample of 47 trauma patients. INTERVENTIONSIntra-assay precision was demonstrated by performance of consecutive analyses of two lactate control solutions (high and low lactate control concentrations) by medical students and physicians. Split sample, simultaneous testing of the portable lactate analyzer was then performed on 66 whole blood specimens from a convenience sample of 47 trauma patients admitted to an urban Level 1 trauma center over 4 mos. Samples were tested simultaneously tested on two portable lactate analyzers and a reference Instrument. MEASUREMENTS AND MAIN RESULTSAcceptable intra-assay precision was achieved. Regression analysis for two test instruments demonstrated a slope of 0.920, an intercept of 0.323, an r of .982, and an SEM of 0.496. Regression analysis for test instrument "A" vs. the reference instrument showed a slope of 0.861, an intercept of 0.209, an r of .977, and an SEM of 0.598. Regression analysis for test instrument "B" vs. the reference instrument demonstrated a slope of 0.929, an intercept of -0.095, an r of .983, and an SEM of 0.506. CONCLUSIONSGood correlation with a low SEM was obtained over a wide range of clinically relevant lactate values. Use of point of care lactate analysis will decrease analytic time, making an important diagnostic parameter immediately available in the critical care setting. (Crit Care Med 1998; 26:1523-1528